The CAT-Tox (L) Assay: A Sensitive and Specific Measure of Stress-Induced Transcription in Transformed Human Liver Cells

Identifying and measuring the molecular mechanisms of toxicity is an important goal in hazard assessment. We have developed an assay in transformed human liver cells to simultaneously measure the transcriptional responses of 14 stress promoter- or response element-chloramphenicol acetyl transferase (CAT) fusion constructs that are stably integrated into the HepG2 cell line. This assay can measure a wide spectrum of stresses, both toxic and nontoxic, such as protein and protein biosynthesis perturbations, DNA damage, heavy metals, and planar aromatic hydrocarbons. We found that each promoter or response element can be induced by one or more of four chemicals that were tested in the assay. These results have been interpreted in light of the current models of action for each compound. The responses of this assay system can distinguish among compounds that are closely related in their structure and have been shown previously to elicit similar biological activities in simple assay systems. We have designated this technique the CAT-Tox (L)iver assay. It measures a broad range of cellular stresses and toxicants at levels that were comparable to or below those of established methods. The induction profiles generated using the CAT-Tox (L) assay can help to elucidate the molecular mechanisms by which chemicals exert their actions on human cells. These profiles can be indicative of both toxic and nontoxic processes that are occurring in the cell. We propose that this cellular stress assay can serve as a screen for a variety of substances at the molecular level.